Use of hexokinase-2 in detection of rare tumor cells in body fluid sample and kit

Abstract

The present application relates to use of hexokinase-2 and a kit for detecting rare tumor cells in body fluid sample. The present application, based on a principle of abnormal tumor cell energy metabolism, conducts a vitro detection by using a glycolysis marker hexokinase-2 (HK2) as a marker, and assists localization and selection of the rare tumor cells by an addressable microporous array chip or a glass sheet, so as to implement a detection on rare tumor cells having high glycolysis activity in a cancer patient's body fluid sample. The detection method of present application allows to detect rare tumor cells having high glycolysis activity in the human body fluid sample, especially detect circulating tumor cells of an interstitial origin tumor and the circulating tumor cells having epithelial-interstitial transformation in epithelial origin tumors, thereby covering the shortage of traditional detection against the circulating tumor cells based on an epithelial marker, and providing a technical basis for better application of cancer liquid biopsy in clinic.

What is claimed is: 
     
       1. A method for detecting circulating tumor cells in a human body fluid sample, comprising:
 incubating the human body fluid sample with a fluorescein labeled hexokinase-2 (HK2) antibody, an antibody specific to a leukocyte marker, and a cell nucleus stain; 
 measuring the level of fluorescein labeled HK2 antibody bound to cells in the human body fluid sample that are stained by the cell nucleus stain, and are not bound to the antibody specific to the leukocyte marker; and 
 identifying cells that are bound to a level of fluorescein labeled HK2 antibody higher than the mean plus five times of standard deviation of reference HK2 antibody levels bound to leukocytes as circulating tumor cells. 
 
     
     
       2. The method according to  claim 1 , wherein the fluorescein labeled HK2 antibody is a fluorescein labeled HK2 antibody, or a HK2 antibody bound to a fluorescein labeled secondary antibody targeting the HK2 antibody. 
     
     
       3. The method according to  claim 1 , wherein the leukocyte marker is cell membrane surface protein CD45. 
     
     
       4. The method according to  claim 1 , wherein the human body fluid sample is selected from the group consisting of blood, pleural effusion, cerebrospinal fluid and urine.